64 MARCH 2018 • WORLD AQUACULTURE • WWW.WAS.ORG Work was done to produce offspring and determine the physiological usefulness of juveniles obtained with the use of cryopreserved sperm (Bogatyreva 2010, Krasilnikova 2015). Fish obtained using thawed sperm had the best survival rate, growth rate, fertility, physiological and biochemical characteristics, compared with fish produced using traditional methods. Years of research to develop methods of cryopreservation of fish eggs in southern Russia has led to the development of a cryoprotectant with “enveloping action,” the basic component of which is a mixture of unrefined vegetable and animal oils containing linoleic, linolenic acids and phosphatides, which are glycerol esters of fatty acids and a substituted phosphoric acid. Comparative evaluation of a standard treatment with this new penetrating cryoprotectant showed that, after freezing in liquid nitrogen, oocytes of the sterlet, Russian sturgeon and Beluga sturgeon were capable of fertilization (20-45 percent). Ongoing collaborative work between the Southern Scientific Center of the Russian Academy of Sciences and Astrakhan State Technical University has led to the creation of a cryobank, whose activities are fundamentally different from existing experimental collections. The basis of these activities has led to the accumulation, conservation and use of genetic material for the shortfall of broodfish and the correction of existing technologies of artificial reproduction of rare and endangered fish species of the Volga-Caspian Sea and Azov-Black Sea basins. The creation of a low-temperature bank will: 1) Keep genetic information of rare and endangered species at the temperature of liquid nitrogen for decades without loss of genetic information. 2) Transport genetic material into the areas with a disappearance or sharp reduction in fish populations for species restoration or to provide genetic material according to the needs of customers; 3) Provide opportunities for breeding and genetic work on fish farms. 4) Create a fairly complete genetic collection of different species of hydrobionts for the subsequent full restoration of hydrobionts of the basin and of ecosystems; 5) Reduce the area used for holding broodfish, thereby improving the economic efficiency of artificial reproduction and commodity fish cultivation. The laboratory has all necessary equipment for the cryobank: cryostorage for long-term storage of sperm samples; for experimental works and transportation of gametes and liquid nitrogen; a programmable freezer; a personal computer; a microscope equipped with a video-ocular for visual determination of sperm quality; fish tanks for the works to produce offspring; and an installation to determine the behavioral responses of juvenile fish. Material will be collected from the hatcheries of the Astrakhan, Volgograd and Rostov regions of Russia, enabling the exchange of genetic material in the southern federal district of Russia. Every year the work of freezing and accumulation of sperm samples of various fish species is carried out. All data about the broodfish from which the sperm is taken is entered into a database, which is used to choose the sample for fertilization of the spawn. Systematic quality control of semen stored in the repository for 1-3 years is implemented. During this time the sperm does not lose its capacity to fertilize eggs. The development of the Southern Scientific Center of the Russian Academy of Sciences and Astrakhan State Technical University in low-temperature sperm cryopreservation of valuable fish species allows for long-term storage of gametes and their use for artificial fertilization. Acknowledgements This article was prepared as part of the state task of the Federal research center of the Southern scientific center of the Russian Academy of Sciences for 2018, No. 01201354245 state registration of the project using a unique modular installation — complex No. 73602 and Bioresources collection of rare and endangered species of fish of the Federal research center of the Southern scientific center of the Russian Academy of Sciences. Work was performed under the Federal Program of research and development on priority directions of scientific-technological complex of Russia for 2014-2020 on the specific project “Development of technical equipment, biotechnology, growing unconventional species of fish and invertebrates for the progress of aquaculture NorthWestern and southern Federal districts of Russia” (agreement No. 14.607.21.0163 on 03.10.2016, at the unique identifier applied research (project) RFMEFI60716X0163), and Grant of President of Russian Federation for state support of young Russian scientists MK-115.2017.11. Notes Aleksandra A. Krasilnikova and Elena N. Ponomareva, Southern Scientific Centre of the Russian Academy of Sciences, 41 Chekhov Street, 344006, Rostov-on-Don, Russia Email: alexandra.kras@yandex.ru and Astrakhan State Technical University, 16 Tatischeva str., Astrakhan, 414056, Russia. Andrey M. Tikhomirov, Astrakhan State Technical University, 16 Tatischeva str., Astrakhan, 414056, Russia. References Bogatyreva, M.M. 2010. Optimization of methods of semen cryopreservation to preserve gene pool of sturgeons. Ph.D. Thesis: Russia, Astrakhan, Astrakhan State Technical University: 126 p. Krasilnikova, A.A. and A.M. Tikhomirov. 2014a. Alternative methods of preparation of fish sperm to freeze at ultra-high values of cooling rate. Vestnik of Astrakhan State Technical University. Series Fisheries 2:72-78. Krasilnikova, A.A. and A.M. Tikhomirov. 2014b.The volume of the frozen sample as one of the factors of survival of spermatozoa of sturgeon fish species in the cryopreservation. Natural Sciences 2(47):62-69. Krasilnikova, A.A. 2015. Improving the process of cryopreservation of the reproductive cells of the male fish. Ph.D. Thesis: Russia, Astrakhan, Astrakhan State Technical University,149 p. Krasilnikova, A.A. and A.M. Tikhomirov. 2015. Correlation of volumes of intracellular fluid, sperm and endocellulases protector in cryoprotective media for sturgeon. Natural Sciences 3(52):96102. Tikhomirov, A.M. and E.N. Ponomareva. 2008. Electrical stimulation of the membranes of sperm of Russian sturgeon facilitates the penetration of cryoprotectants into cells. Biophysics of the living cell. Conservation of Genetic Resources 9:129-130.
RkJQdWJsaXNoZXIy MjExNDY=